在金纳米粒子和氧化石墨烯上使用立足点介导的链置换扩增法基于电化学适体测定蛋白质酪氨酸激酶-7,Microchimica Acta

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在金纳米粒子和氧化石墨烯上使用立足点介导的链置换扩增法基于电化学适体测定蛋白质酪氨酸激酶-7
Microchimica Acta ( IF 5.3 ) Pub Date : 2019-10-26 , DOI: 10.1007/s00604-019-3849-z
Zongbing Li ₁ , Zhengkun Zhou ₁ , Ning Xue ₁ , Shujie Wu ₁ , Xiangmin Miao ₁

Affiliation

描述了一种用于超灵敏测定蛋白质酪氨酸激酶 7 (PTK7) 的电化学方法。它基于 (a) 使用带正电荷的金纳米粒子 (AuNPs) 和带负电荷的氧化石墨烯 (GO)，以及 (b) 立足点介导的链置换扩增。包含 sgc8 适体的发夹探针 2 (HP2) 用于修饰玻碳电极 (GCE)。它的发夹结构在 PTK7 存在下打开，形成 PTK7-HP2 复合物。HP2 的暴露部分与固定在 AuNP 和 GO 修饰的 GCE 上的发夹探针 1 (HP1) 部分杂交。添加用氧化还原探针亚甲蓝 (MB-HP3) 标记的发夹探针 3 后，由于 HP1 与 MB-HP3 的互补杂交，会发生立足点介导的链置换。这导致 PTK7-HP2 释放到溶液中并使其可用于下一个反应。在最佳条件下，PTK7 可以通过伏安法（通常在 -0.18 V 下进行）进行量化，检测限为 1.8 fM。由于使用了适体，该测定对 PTK7 具有高选择性。成功应用于恶性黑色素瘤A375细胞碎片中PTK7的测定。图形摘要基于金纳米粒子和氧化石墨烯上的立足点介导的链置换反应放大，用于超灵敏测定蛋白酪氨酸激酶 7 (PTK7) 的无酶电化学传感器的示意图。

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Electrochemical aptamer-based determination of protein tyrosine kinase-7 using toehold-mediated strand displacement amplification on gold nanoparticles and graphene oxide

An electrochemical method is described for ultrasensitive determination of protein tyrosine kinase-7 (PTK7). It is based on (a) the use of positively charged gold nanoparticles (AuNPs) and negatively charged graphene oxide (GO), and (b) of toehold-mediated strand displacement amplification. A hairpin probe 2 (HP2) containing the sgc8 aptamer was used to modify a glassy carbon electrode (GCE). Its hairpin structure is opened in the presence of PTK7 to form the PTK7-HP2 complex. The exposed part of HP2 partly hybridizes with hairpin probe 1 (HP1) that was immobilizing on the AuNPs and GO modified GCE. On addition of the hairpin probe 3 that was labeled with the redox probe Methylene Blue (MB-HP3), toehold-mediated strand displacement occurs due to complementary hybridization of HP1 with MB-HP3. This causes the release of PTK7-HP2 into the solution and makes it available for the next reaction. Under optimal conditions, PTK7 can be quantified by voltammetry (typically performed at −0.18 V) with a detection limit of 1.8 fM. The assay possesses high selectivity for PTK7 due to the employment of the aptamer. It was successfully applied to the determination of PTK7 in the debris of malignant melanoma A375 cells. Graphical abstract Schematic representation of the enzyme-free electrochemical sensor for ultrasensitive determination of protein tyrosine kinase-7 (PTK7) based on the toehold-mediated strand displacement reaction amplification on gold nanoparticles and graphene oxide. Schematic representation of the enzyme-free electrochemical sensor for ultrasensitive determination of protein tyrosine kinase-7 (PTK7) based on the toehold-mediated strand displacement reaction amplification on gold nanoparticles and graphene oxide.

更新日期：2019-10-26

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